An Optimized Genomic DNA Isolation Protocol for the High-value Medicinal herb Allium stracheyi Baker for Molecular Marker-based Analysis

Ashish Kumar¹, Yogita Bisht¹, Arun Kumar Jugran¹and Surendra Singh Bargali^2*

¹G. B. Pant National Institute of Himalayan Environment, Garhwal Regional Centre, Upper Bhaktiyana, Srinagar Garhwal, Uttarakhand, India

²Department of Botany, D.S.B. Campus, Kumaun University, Nainital, Uttarakhand, India

Corresponding Author E-mail:surendrakiran@rediffmail.com

Abstract:

Obtaining intact genomic DNA of adequate purity from high- altitude Himalayan medicinal plants is often difficult because their tissues contain substantial amounts of phenolic compounds, polysaccharides and other metabolites that can interfere with DNA purification and subsequent molecular analyses. In this study, we optimized a modified CTAB procedure for efficient recovery of genomic DNA from the leaves of Allium stracheyi Baker. The optimized system incorporated 15% polyvinylpyrrolidone (PVP) and 2% β-mercaptoethanol (β-ME), together with a 5 M NaCl-assisted precipitation step to reduce polysaccharide and secondary-metabolite contamination. The quality of DNA was further validated through PCR amplification using ISSR and chloroplast SSR molecular markers, producing clear and reproducible banding patterns. The developed protocol is reliable and suitable for molecular studies, including genetic diversity and conservation research in A. stracheyi and other medicinal plant species in secondary metabolites.

Keywords:

Allium stracheyi; Genomic DNA isolation; Himalayan medicinal plant; Molecular marker; Polymerase Chain Reaction

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